間接検定調査牛における脂肪交雑関連遺伝子SNP多型の効果の推定

• 資料種別: 博士論文
• タイトル: 間接検定調査牛における脂肪交雑関連遺伝子SNP多型の効果の推定
• 著者・編者: 佐々木, 整輝
• 出版事項: 新潟大学
• 出版年月日等: 2014-03-24
• 出版年（W3CDTF）: 2014-03-24
• 並列タイトル等: 間接検定調査牛における脂肪交雑関連遺伝子SNP多型の効果の推定
• 授与機関名: 新潟大学
• 授与年月日: 2014-03-24
• 授与年月日（W3CDTF）: 2014-03-24
• 報告番号: 甲第3936号
• 学位: 博士（農学）
• 博論授与番号: 甲第3936号
• 本文の言語コード: jpn
• 一般注記: Marbling defined by the amount and distribution of intramuscular fat is an economically important trait of beef cattle in Japan. While competition with imported meat is intensifying, it will become increasingly important for meat production of Japan to improve the quality and reduce the cost of production. In improvement of marbling, genetic evaluation calculated by BLUP method (i. e. estimated breeding value: EBV) have been used. In recent years, to perform genetic evaluation more accurately, a lot of studies begin to try identifying the genes that responsible for economical trait. Previous studies have reported, five single nucleotide polymorphisms (SNPs) in endothelial differentiation sphingolipid G-protein-coupled receptor 1 (EDG1), titin(TTN), akirin 2(AKIRIN2), ribosomal protein L27a(RPL27A) were associated with marbling in Japanese Black beef cattle population of the Oita prefecture (Yamada et al. 2009a; 2009b, 2009c; 2009d). The SNPs were reported to be located in the promoter region of EDG1 (g. 1471620G>T), the untranslated region of EDG1 (c. -312A>G), the promoter region of TTN (g. 231054C>T), the 3’ untranslated region ofAKIRIN2 and the promoter region of RPL27A (g. 3109537C>T) respectively. However, the results of these associations were studied obtained only the population which consists of limited strain in a part of area, thus it is important to confirm usefulness of these SNPs for establishment of breeding program of whole Japanese black cattle populations. The objectives of this study were to estimate an effect of the five SNP genotypes on marbling more exactly and to analyze association of the SNP genotypes with the carcass traits other than the BMS, by utilizing a large population consisting of various strains. For this purpose, our present study used large population of 1, 351 Japanese black steers, in which the environmental factors were controlled and various strains were included in the Livestock Improvement Association of Japan. For the necessary to genotype the five SNPs for a large number of DNA samples, in the second chapter, we developed a new protocol, which was named "2 STEP-MULTIPLE-PCR-SSP method", by modifying the Sequence-Specific Primer Cycle Elongation (SSPCE) method. The main feature of this method is that it is performed in two steps. In the first step, the SNP-containing target region which spanned 100～300 bp were amplified by PCR, In the second step, allele specific reaction was carried out to detect genotype. When the series of these procedures were performed with the optimized conditions, in the 1st step, 5 different PCR products from the five SNPs were observed definitely, and in the 2nd step, allele specific reaction occurred in each SNP sites correctly. Furthermore, genotyping results of the 2STEP-MULTIPLE-PCR-SSP were in complete agreement with genotypes of previous study. This method should be useful for a several SNPs typing a number of samples, because it requires neither particular equipment nor expensive kit, and it also is easy to optimize a condition of reaction.; In the third Chapter, 1, 351 Japanese black steers population that took progeny test in the Livestock Improvement Association of Japan were genotyped with 2STEP-MULTIPLE-PCR-SSP method, and associations of these 5 SNPs with beef marbling standard number (BMS) were investigated. The raw data of recorded BMS should contain environmental variances, so that it is impossible to exactly predict allele substitution effect using its raw data. Thus, we used estimated breeding value (EBV) which were calculated by BLUP method to analyze association between the SNPs and BMS, as a result of association analysis between EBV of BMS and the SNPs, statistically significant differences were detected on c.-312A>G and g. 231054C>T in EDG1 and TTN genes, respectively. But no significant differences were detected on g. 1471620G>T in EDG1, c. 188G>A in AKIRIN2, and g.3109537C>T in RPL2 7A. Differences between least squares means for genotypes within a SNP were examined by Tukey-Kramer s test. As a result, GG homozygotes showed significantly higher value than AA homozygotes in EDG1:c.-312A>G. The differences between AA and AG genotypes were 0. 13 and the differences between AG and GG genotypes were 0. 11.TT homozygotes also exhibited significantly higher value than the other genotypes in TTN:g.231054C>T. The differences between CC and TC genotypes were 0.06 and the differences between CT and TT genotypes were 0.40. In the other hand, no inter-allelic interaction between EDG1 and TTN was detected. Thus, I concluded that the SNPs in EDG1:C.-312A>G and TTN:g.231054C>T may be regarded as useful markers for breeding program such as selection or evaluation. Whereas the two SNPs are useful for selection or evaluation for BMS, attention should be paid to other trait such as carcass weight, feed efficiency and so on. Consequently, in the fourth chapter, we investigated whether EDG1:c.-312A>G and TTN:g.231054C>T affect other traits. The association of the EDG1:c.-312A>G and TTN;g.231054C>T with the subcutaneous fat start weight(SWT), end weight(ET), start height(SH), end height(EWI-I), daily gain, subcutaneous fat thickness (SFT), carcass weight (CWT), rib eye area (REA), and rib thickness (RT), were studied. The breeding values for each trait were predicted by BLUP in the same way as the BMS, prior to the analysis of association between the two SNPs and these traits. In the case of EDG1:c.-312A>G, statistically significant differences were detected in EWT, DG, CWT, RT and SFT, but not in SWT, SH and EH. In these traits, G allele of EDG1:c.-312A>G was more favorable than A allele for meat production. In the case of TTN:g. 231054C>T, statistically significant differences were detected in DG and RT, and T allele of TTN:g.231054C>T was more favorable than C allele for meat production. EDG1 is involved in angiogenesis in terms of function, therefore active expression of this gene result in the development of mature vessels, in which nutrient are delivered. On the other hand, TTN encodes a protein that is responsible for passive elasticity of muscle, active expression of this gene leads to the muscle development. The fact of this function suggested that EDG1 and TTN exert a function in physiological or anatomical milieux surrounding adipocyte-linage cells rather than intramuscular adipocyte-linage cells. The milieux were thought to promote proliferation, differentiation or maturation of adipocyte-linage cells by controlling intramuscular vascularization and structural integrity of the sarcomere. This study demonstrated that G allele of EDGKc.-312A〉G and T allele TTN:g. 231054C>T revealed favorable effects on both of beef quality and productivity, suggesting that the two SNPs might be able to apply to breeding program for Japanese Black beef cattle.; 学位の種類: 博士（農学）. 報告番号: 甲第3936号. 学位記番号: 新大院博（農）甲第139号. 学位授与年月日: 平成26年3月24日; 新大院博（農）甲第139号; 開始ページ : 1; 終了ページ : 80
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• 公開開始日（W3CDTF）: 2015-03-03
• 受理日（W3CDTF）: 2015-02-03T05:25:05+09:00
• 作成日（W3CDTF）: 2019-08-05
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